Detergent compositions containing bacillus agaradhaerens mannanase and methods of use thereof

Abstract

The present compositions and methods relate to an endo-beta-mannanase cloned from Bacillus agaradhaerens, polynucleotides encoding the endo-beta-mannanase, and methods of use thereof. Formulations containing the endo-beta-mannanase are highly suitable for use as detergents.

Description

[0001] priority

[0002] This patent application claims priority to International Application No. PCT / CN2011 / 073525 filed on April 29, 2011, which is hereby incorporated by reference in its entirety. technical field

[0003] The compositions and methods of the present invention relate to endo-β-mannanases cloned from Bacillus agaricus, polynucleotides encoding endo-β-mannanases, and methods of their use. Formulations comprising endo-beta-mannanase are very suitable for use as detergents. Background technique

[0004] Current laundry detergent and fabric care compositions include ingredients such as surfactants, enzymes (proteases, amylases, mannanases and / or cellulases), bleaches, builder systems, suds suppressors Complex combinations of active ingredients such as , stain suspending agents, soil release agents, optical brighteners, softeners, dispersants, dye transfer inhibiting compounds, abrasives, bactericides and fragrances.

[0005] Mannanases, including endo-beta-man...

Examples

example

[0246] The following examples are provided to demonstrate and illustrate certain preferred embodiments and aspects of the invention and should not be construed as limiting.

[0247] In the following experimental disclosures, the following abbreviations are used: M (mole / liter), mM (millimol / liter), μM (micromoles / liter), nM (nanomoles / liter), mol (mole), mmol (mmol ), μmol (micromole), nmol (nanomole), g and gm (gram), mg (milligram), μg (microgram), pg (picogram), L (liter), ml and mL (milliliter), μl and μL (microliter), cm (centimeter), mm (millimeter), μm (micrometer), nm (nanometer), U (unit), MW (molecular weight), s (second), min (minute), h (hour ), ℃ (Celsius), QS (sufficient), ND (not performed), rpm (rev / min), H 2 O (water), dH 2 O (deionized water), HCl (hydrochloric acid), aa (amino acid), bp (base pair), kb (kilobase pair), kD (kilodalton), MgCl 2 (magnesium chloride), NaCl (sodium chloride), Ca (calcium), Mg (magnesium), HEPES (4-(2-hydroxyethyl)-1-piperazine...

example 1

[0249] Cloning of Glycosyl Hydrolase Bag Man1 from Bacillus agaricus

[0250] Bacillus agaricus was selected as a potential source of a variety of glycosyl hydrolases and other enzymes that can be used in industrial applications. Genomic DNA for sequencing was obtained as follows: first, a strain of Bacillus agaricus was placed on a GAM agar plate at 30°C (Jones et al., IJSEM, 55:1711-1714, 2005 (Jones et al., "International Journal of Systematic and Evolutionary Microbiology, Vol. 55, pp. 1711-1714, 2005)) Incubation for 24 hours. Cellular material was scraped from the plates and used to prepare genomic DNA with the aid of the ZF Fungal / Bacterial DNA Miniprep Kit from Zymo (Cat# D6005). The genomic DNA was used for sequencing and used to amplify the Bag Man1 gene for expression cloning. use Sequencing by synthesis (SBS) technology (www.baseclear.com / sequencing / illumina-sequencing) Whole-genome sequencing of Bacillus agaricus viscosus. Genome sequencing and sequence dat...

example 2

[0256] Expression of Glycosyl Hydrolase (Bag Man1) from Bacillus agaricus

[0257] The Bag Man1 gene was amplified by PCR from Bacillus agaricus gluei genomic DNA using the following primers: Primer 1 (BssHII) 5'-TGAGCGCGCA GGCTGCTGGA AAAATGACAA ACTATGAATC AGAGGT-3' (SEQ ID NO: 8), and Primer 2 (BamHI) 5 '-TGTGGATCCT TACTCTAACG GTACGTCTTC CTTAT-3' (SEQ ID NO: 9). The amplified Bag Man1 gene was cloned into the expression vector p2JM by BssHII / BamHI double enzyme digestion and ligation. The Bacillus subtilis expression vector p2JM103BBI (Vogtentanz, Protein Expr Purif, 55:40-52, 2007) was digested with restriction enzymes BssHII and BamHI (Vogtentanz, "Protein Expression and Purification", Vol. 55, pp. 40-52, 2007)). A DNA fragment free of the BCE103-BBI fusion gene sequence was isolated and used as the expression backbone. This DNA fragment was ligated with the PCR amplified gene encoding the BagManl mature protein, which resulted in the addition of three codons between t...

[0001] priority

[0002] This patent application claims priority to International Application No. PCT / CN2011 / 073525 filed on April 29, 2011, which is hereby incorporated by reference in its entirety. technical field

[0003] The compositions and methods of the present invention relate to endo-β-mannanases cloned from Bacillus agaricus, polynucleotides encoding endo-β-mannanases, and methods of their use. Formulations comprising endo-beta-mannanase are very suitable for use as detergents. Background technique

[0004] Current laundry detergent and fabric care compositions include ingredients such as surfactants, enzymes (proteases, amylases, mannanases and / or cellulases), bleaches, builder systems, suds suppressors Complex combinations of active ingredients such as , stain suspending agents, soil release agents, optical brighteners, softeners, dispersants, dye transfer inhibiting compounds, abrasives, bactericides and fragrances.

[0005] Mannanases, including endo-beta-man...

Examples

example

[0246] The following examples are provided to demonstrate and illustrate certain preferred embodiments and aspects of the invention and should not be construed as limiting.

[0247] In the following experimental disclosures, the following abbreviations are used: M (mole / liter), mM (millimol / liter), μM (micromoles / liter), nM (nanomoles / liter), mol (mole), mmol (mmol ), μmol (micromole), nmol (nanomole), g and gm (gram), mg (milligram), μg (microgram), pg (picogram), L (liter), ml and mL (milliliter), μl and μL (microliter), cm (centimeter), mm (millimeter), μm (micrometer), nm (nanometer), U (unit), MW (molecular weight), s (second), min (minute), h (hour ), ℃ (Celsius), QS (sufficient), ND (not performed), rpm (rev / min), H 2 O (water), dH 2 O (deionized water), HCl (hydrochloric acid), aa (amino acid), bp (base pair), kb (kilobase pair), kD (kilodalton), MgCl 2 (magnesium chloride), NaCl (sodium chloride), Ca (calcium), Mg (magnesium), HEPES (4-(2-hydroxyethyl)-1-piperazine...

example 1

[0249] Cloning of Glycosyl Hydrolase Bag Man1 from Bacillus agaricus

[0250] Bacillus agaricus was selected as a potential source of a variety of glycosyl hydrolases and other enzymes that can be used in industrial applications. Genomic DNA for sequencing was obtained as follows: first, a strain of Bacillus agaricus was placed on a GAM agar plate at 30°C (Jones et al., IJSEM, 55:1711-1714, 2005 (Jones et al., "International Journal of Systematic and Evolutionary Microbiology, Vol. 55, pp. 1711-1714, 2005)) Incubation for 24 hours. Cellular material was scraped from the plates and used to prepare genomic DNA with the aid of the ZF Fungal / Bacterial DNA Miniprep Kit from Zymo (Cat# D6005). The genomic DNA was used for sequencing and used to amplify the Bag Man1 gene for expression cloning. use Sequencing by synthesis (SBS) technology (www.baseclear.com / sequencing / illumina-sequencing) Whole-genome sequencing of Bacillus agaricus viscosus. Genome sequencing and sequence dat...

example 2

[0256] Expression of Glycosyl Hydrolase (Bag Man1) from Bacillus agaricus

[0257] The Bag Man1 gene was amplified by PCR from Bacillus agaricus gluei genomic DNA using the following primers: Primer 1 (BssHII) 5'-TGAGCGCGCA GGCTGCTGGA AAAATGACAA ACTATGAATC AGAGGT-3' (SEQ ID NO: 8), and Primer 2 (BamHI) 5 '-TGTGGATCCT TACTCTAACG GTACGTCTTC CTTAT-3' (SEQ ID NO: 9). The amplified Bag Man1 gene was cloned into the expression vector p2JM by BssHII / BamHI double enzyme digestion and ligation. The Bacillus subtilis expression vector p2JM103BBI (Vogtentanz, Protein Expr Purif, 55:40-52, 2007) was digested with restriction enzymes BssHII and BamHI (Vogtentanz, "Protein Expression and Purification", Vol. 55, pp. 40-52, 2007)). A DNA fragment free of the BCE103-BBI fusion gene sequence was isolated and used as the expression backbone. This DNA fragment was ligated with the PCR amplified gene encoding the BagManl mature protein, which resulted in the addition of three codons between t...

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